Therapeutic effects of Sheng Xue Fang in a cyclophosphamide-induced anaemia mouse model

Sheng Xue Fang (SXF) has been used to treat anaemia for decades with good efficacy. To study the effect and possible mechanism of SXF to restore haematopoietic function. Balb/c mice (10 per/group, half male, half female) were treated with SXF (three dose groups, 8.5, 17, and 22.1 g/kg) by gavage for 14 days, and cyclophosphamide (80 mg/kg) was injected on days 10–12. Only injection of cyclophosphamide (negative control) or physiological saline (blank control) were included as controls. The spleen and femur were processed for histopathology. Active components and the target of SXF were screened. The target was used for gene enrichment and protein-protein interaction (PPI) analysis. Red blood cell relative changes in the SXF group (low: −5.50 ± 1.58%; medium: −11.11 ± 4.15%; high: −8.81 ± 2.67%) and relative negative control (26.21 ± 2.51%) significantly increased (all <i>p</i> &lt; 0.01) in female mice. Haemoglobin and red blood cell-specific volume showed the same trend. However, SXF did not have significant effects on male mice. Splenic index in the medium group (4.44 ± 0.46%) relative negative control (3.38 ± 0.10%) significantly improved (<i>p</i> &lt; 0.01) in female mice. Using network pharmacology, 77 active components and 337 targets were screened from SXF. These targets are closely related to the mitogen-activated protein kinase pathway. SXF has good clinical application potential. However, the mechanism requires in-depth research. Our findings are of great significance in anaemia treatment and provide a new perspective for Chinese medicine research.

升血方（Sheng Xue Fang, SXF）在临床用于治疗贫血已有数十年，疗效确切。为探究升血方对造血功能的恢复作用及其潜在机制，本实验采用BALB/c小鼠（每组10只，雌雄各半），通过灌胃给予升血方，设置三个剂量组（8.5、17、22.1 g/kg），连续给药14天；于第10至12天腹腔注射环磷酰胺（80 mg/kg）构建贫血模型。实验设置单纯注射环磷酰胺组（阴性对照组）与生理盐水组（空白对照组）作为对照。采集脾脏与股骨样本进行组织病理学检测，同时筛选升血方的活性成分与作用靶点，并基于靶点进行基因富集分析及蛋白质相互作用（PPI）网络分析。
结果显示，雌性小鼠中，升血方各剂量组[低剂量组：-5.50±1.58%；中剂量组：-11.11±4.15%；高剂量组：-8.81±2.67%]与阴性对照组（26.21±2.51%）的红细胞相对变化量均显著升高（均p<0.01）；血红蛋白水平与红细胞比容亦呈现相同变化趋势。但升血方对雄性小鼠无显著改善作用。雌性小鼠中，中剂量组的脾脏指数（4.44±0.46%）相较于阴性对照组（3.38±0.10%）显著提升（p<0.01）。
通过网络药理学方法，从升血方中共筛选得到77种活性成分与337个作用靶点，上述靶点与丝裂原活化蛋白激酶（mitogen-activated protein kinase, MAPK）通路密切相关。升血方具有良好的临床应用潜力，但其具体作用机制仍需进一步深入研究。本研究结果对贫血治疗具有重要指导意义，同时为中医药研究提供了全新视角。